Domain-focused CRISPR screen identifies HRI as a fetal hemoglobin regulator in human erythroid cells.
Jeremy D GrevetXianjiang LanNicole HamagamiChristopher R EdwardsLaavanya SankaranarayananXinjun JiSaurabh K BhardwajCarolyne J FaceDavid F PosoccoOsheiza AbdulmalikCheryl A KellerBelinda M GiardineSimone SidoliBen A GarciaStella T ChouStephen A LiebhaberRoss C HardisonJunwei ShiGerd A BlobelPublished in: Science (New York, N.Y.) (2018)
Increasing fetal hemoglobin (HbF) levels in adult red blood cells provides clinical benefit to patients with sickle cell disease and some forms of β-thalassemia. To identify potentially druggable HbF regulators in adult human erythroid cells, we employed a protein kinase domain-focused CRISPR-Cas9-based genetic screen with a newly optimized single-guide RNA scaffold. The screen uncovered the heme-regulated inhibitor HRI (also known as EIF2AK1), an erythroid-specific kinase that controls protein translation, as an HbF repressor. HRI depletion markedly increased HbF production in a specific manner and reduced sickling in cultured erythroid cells. Diminished expression of the HbF repressor BCL11A accounted in large part for the effects of HRI depletion. Taken together, these results suggest HRI as a potential therapeutic target for hemoglobinopathies.
Keyphrases
- induced apoptosis
- crispr cas
- endothelial cells
- red blood cell
- cell cycle arrest
- high throughput
- genome editing
- protein kinase
- transcription factor
- endoplasmic reticulum stress
- end stage renal disease
- signaling pathway
- chronic kidney disease
- gene expression
- small molecule
- peritoneal dialysis
- young adults
- risk assessment
- amino acid
- protein protein